Open-cellular metal implant design and fabrication for biomechanical compatibility with bone using electron beam melting.

Implant history extends more than 4000 years in antiquity, with biocompatible alloy implants extending over only 70 years. Over the past several decades, total hip and knee replacements of Ti-6Al-4V and Co-Cr-Mo alloys have exhibited post implantation life spans extending over 15 years; limited by infection, loosening, stress-shielding-related bone resorption and other mechanical failures. With the advent of additive manufacturing technologies, such as electron beam melting (EBM) over the past decade, personalized, patient-specific; porous (open-cellular) implant components can be manufactured, and the integration of chemical, biological and mechanical methods is able to optimize strategies for improving long-term clinical outcomes. This review outlines these strategies, which include enhanced osseointegration and vascularization prospects, and provides some evidence for, and examples of, clinical trials representative of millions of implant surgeries world-wide.

Interplay between self-assembled structure of bone morphogenetic protein-2 (BMP-2) and osteoblast functions in three-dimensional titanium alloy scaffolds: Stimulation of osteogenic activity.

Three-dimensional cellular scaffolds are receiving significant attention in bone tissue engineering to treat segmental bone defects. However, there are indications of lack of significant osteoinductive ability of three-dimensional cellular scaffolds. In this regard, the objective of the study is to elucidate the interplay between bone morphogenetic protein (BMP-2) and osteoblast functions on 3D mesh structures with different porosities and pore size that were fabricated by electron beam melting. Self-assembled dendritic microstructure with interconnected cellular-type morphology of BMP-2 on 3D scaffolds stimulated osteoblast functions including adhesion, proliferation, and mineralization, with prominent effect on 2-mm mesh. Furthermore, immunofluorescence studies demonstrated higher density and viability of osteoblasts on lower porosity mesh structure (2 mm) as compared to 3- and 4-mm mesh structures. Enhanced filopodia cellular extensions with extensive cell spreading was observed on BMP-2 treated mesh structures, a behavior that is attributed to the unique self-assembled structure of BMP-2 that effectively communicates with the cells. The study underscores the potential of BMP-2 in imparting osteoinductive capability to the 3D printed scaffolds.

Influence of cell shape on mechanical properties of Ti-6Al-4V meshes fabricated by electron beam melting method.

Ti-6Al-4V reticulated meshes with different elements (cubic, G7 and rhombic dodecahedron) in Materialise software were fabricated by additive manufacturing using the electron beam melting (EBM) method, and the effects of cell shape on the mechanical properties of these samples were studied. The results showed that these cellular structures with porosities of 88-58% had compressive strength and elastic modulus in the range 10-300MPa and 0.5-15GPa, respectively. The compressive strength and deformation behavior of these meshes were determined by the coupling of the buckling and bending deformation of struts. Meshes that were dominated by buckling deformation showed relatively high collapse strength and were prone to exhibit brittle characteristics in their stress-strain curves. For meshes dominated by bending deformation, the elastic deformation corresponded well to the Gibson-Ashby model. By enhancing the effect of bending deformation, the stress-strain curve characteristics can change from brittle to ductile (the smooth plateau area). Therefore, Ti-6Al-4V cellular solids with high strength, low modulus and desirable deformation behavior could be fabricated through the cell shape design using the EBM technique.

Compression deformation behavior of Ti-6Al-4V alloy with cellular structures fabricated by electron beam melting.

Ti-6Al-4V alloy with two kinds of open cellular structures of stochastic foam and reticulated mesh was fabricated by additive manufacturing (AM) using electron beam melting (EBM), and microstructure and mechanical properties of these samples with high porosity in the range of 62%∼92% were investigated. Optical observations found that the cell struts and ligaments consist of primary α' martensite. These cellular structures have comparable compressive strength (4∼113 MPa) and elastic modulus (0.2∼6.3 GPa) to those of trabecular and cortical bone. The regular mesh structures exhibit higher specific strength than other reported metallic foams under the condition of identical specific stiffness. During the compression, these EBM samples have a brittle response and undergo catastrophic failure after forming crush band at their peak loading. These bands have identical angle of ∼45° with compression axis for the regular reticulated meshes and such failure phenomenon was explained by considering the cell structure. Relative strength and density follow a linear relation as described by the well-known Gibson-Ashby model but its exponential factor is ∼2.2, which is relative higher than the idea value of 1.5 derived from the model.

In vivo corrosion, tumor outcome, and microarray gene expression for two types of muscle-implanted tungsten alloys.

Tungsten alloys are composed of tungsten microparticles embedded in a solid matrix of transition metals such as nickel, cobalt, or iron. To understand the toxicology of these alloys, male F344 rats were intramuscularly implanted with pellets of tungsten/nickel/cobalt, tungsten/nickel/iron, or pure tungsten, with tantalum pellets as a negative control. Between 6 and 12 months, aggressive rhabdomyosarcomas formed around tungsten/nickel/cobalt pellets, while those of tungsten/nickel/iron or pure tungsten did not cause cancers. Electron microscopy showed a progressive corrosion of the matrix phase of tungsten/nickel/cobalt pellets over 6 months, accompanied by high urinary concentrations of nickel and cobalt. In contrast, non-carcinogenic tungsten/nickel/iron pellets were minimally corroded and urinary metals were low; these pellets having developed a surface oxide layer in vivo that may have restricted the mobilization of carcinogenic nickel. Microarray analysis of tumors revealed large changes in gene expression compared with normal muscle, with biological processes involving the cell cycle significantly up-regulated and those involved with muscle development and differentiation significantly down-regulated. Top KEGG pathways disrupted were adherens junction, p53 signaling, and the cell cycle. Chromosomal enrichment analysis of genes showed a highly significant impact at cytoband 7q22 (chromosome 7) which included mouse double minute (MDM2) and cyclin-dependant kinase (CDK4) as well as other genes associated with human sarcomas. In conclusion, the tumorigenic potential of implanted tungsten alloys is related to mobilization of carcinogenic metals nickel and cobalt from corroding pellets, while gene expression changes in the consequent tumors are similar to radiation induced animal sarcomas as well as sporadic human sarcomas.

Microstructure and mechanical properties of open-cellular biomaterials prototypes for total knee replacement implants fabricated by electron beam melting.

Total knee replacement implants consisting of a Co-29Cr-6Mo alloy femoral component and a Ti-6Al-4V tibial component are the basis for the additive manufacturing of novel solid, mesh, and foam monoliths using electron beam melting (EBM). Ti-6Al-4V solid prototype microstructures were primarily α-phase acicular platelets while the mesh and foam structures were characterized by α(')-martensite with some residual α. The Co-29Cr-6Mo containing 0.22% C formed columnar (directional) Cr(23)C(6) carbides spaced ~2 µm in the build direction, while HIP-annealed Co-Cr alloy exhibited an intrinsic stacking fault microstructure. A log-log plot of relative stiffness versus relative density for Ti-6Al-4V and Co-29Cr-6Mo open-cellular mesh and foams resulted in a fitted line with a nearly ideal slope, n = 2.1. A stress shielding design graph constructed from these data permitted mesh and foam implant prototypes to be fabricated for compatible bone stiffness.

Evidence of low-temperature superparamagnetism in Mn3O4 nanoparticle ensembles.

Using ac-susceptibility, dc-magnetization, and transmission electron microscopy, we have investigated the magnetic behavior of Mn(3)O(4) nanoparticle ensembles at temperatures below the paramagnetic-to-ferrimagnetic transition of the title material (T(N) approximately equal 41 K). Our data show no evidence of the complex magnetic ordering exhibited by bulk Mn(3)O(4), or of a magnetic behavior around T(N) that has a dynamic (relaxation) origin. Instead, we find a low-temperature (at approximately 11 K) magnetic anomaly that manifests itself as a peak in the out-of-phase component of the ac-susceptibility. Analysis of the frequency and average-particle-size dependence of the peak temperature demonstrates that this behavior is due to the onset of superparamagnetic relaxation, and not to a previously hinted at spin-glass-like transition. Indeed, the relative peak temperature variation per frequency decade DeltaT/TDeltalog(f) is 0.11, an order of magnitude larger than the value expected for collective spin freezing, but within the range of values observed for superparamagnetic blocking. Furthermore, attempts to fit the frequency f/observation time tau = 1/2pif dependence of the peak temperature by a power law led to parameter values unexpected for a spin-glass transition. On the other hand, a Vogel-Fulcher law tau = tau(0)exp[E(B)/k(B)(T - T(0))]-where E(B) is the energy barrier to magnetization reversal, k(B) is the Boltzmann constant, tau(0) and T(0) are constants related to the attempt frequency and the interparticle interaction strength-correctly describes the peak shift and yields values consistent with the superparamagnetic behavior of a slightly interacting system of nanoparticles. In addition, the peak temperature T is sensitive to minute changes in the average particle size (D), and scales as (T - T(0) is proportional to(D)3, another signature of superparamagnetic relaxation.

Next-generation biomedical implants using additive manufacturing of complex, cellular and functional mesh arrays.

In this paper, we examine prospects for the manufacture of patient-specific biomedical implants replacing hard tissues (bone), particularly knee and hip stems and large bone (femoral) intramedullary rods, using additive manufacturing (AM) by electron beam melting (EBM). Of particular interest is the fabrication of complex functional (biocompatible) mesh arrays. Mesh elements or unit cells can be divided into different regions in order to use different cell designs in different areas of the component to produce various or continually varying (functionally graded) mesh densities. Numerous design elements have been used to fabricate prototypes by AM using EBM of Ti-6Al-4V powders, where the densities have been compared with the elastic (Young) moduli determined by resonant frequency and damping analysis. Density optimization at the bone-implant interface can allow for bone ingrowth and cementless implant components. Computerized tomography (CT) scans of metal (aluminium alloy) foam have also allowed for the building of Ti-6Al-4V foams by embedding the digital-layered scans in computer-aided design or software models for EBM. Variations in mesh complexity and especially strut (or truss) dimensions alter the cooling and solidification rate, which alters the alpha-phase (hexagonal close-packed) microstructure by creating mixtures of alpha/alpha' (martensite) observed by optical and electron metallography. Microindentation hardness measurements are characteristic of these microstructures and microstructure mixtures (alpha/alpha') and sizes.

Microstructure and mechanical behavior of Ti-6Al-4V produced by rapid-layer manufacturing, for biomedical applications.

The microstructure and mechanical behavior of simple product geometries produced by layered manufacturing using the electron beam melting (EBM) process and the selective laser melting (SLM) process are compared with those characteristic of conventional wrought and cast products of Ti-6Al-4V. Microstructures are characterized utilizing optical metallography (OM), scanning electron microscopy (SEM) and transmission electron microscopy (TEM), and included alpha (hcp), beta (bcc) and alpha(') (hcp) martensite phase regimes which give rise to hardness variations ranging from HRC 37 to 57 and tensile strengths ranging from 0.9 to 1.45 GPa. The advantages and disadvantages of layered manufacturing utilizing initial powders in custom building of biomedical components by EBM and SLM in contrast to conventional manufacturing from Ti-6Al-4V wrought bar stock are discussed.

Cytotoxic responses and potential respiratory health effects of carbon and carbonaceous nanoparticulates in the Paso del Norte airshed environment.

We have utilized a range of manufactured or commercial nanoparticulate materials, including surrogate carbon nano-PM along with combustion-generated carbonaceous (soot) nano-PM characteristic of environmental nano- PM (both indoor and outdoor) to investigate and compare their cytotoxic response in vitro with an immortalized human epithelial (lung model) cell line (A549). These have included nano-Ag, Al2O3, TiO2, Fe2O3, ZrO2, Si3N4, chrysotile asbestos, BC, 2 types of MWCNT-aggregate PM (MWCNT-R and MWCNT-N), and high-volume glass fiber collected soots: candle, wood, diesel (truck), tire, and 3-types of natural gas kitchen burner-generated soots: yellow (fuel-rich) flame, low-flow blue flame, and normal flow blue flame soot PM. These carbonaceous nano-PM species can be found in either the indoor and outdoor environments or microenvironments. Two-day and two-week in-vitro cultures of A549 showed cell death (or decreased cell viability) for all nanoparticulate materials, but especially significant for all but the TiO2 and candle, wood, and diesel PM. The natural gas kitchen burner combustion PM cell death response was characteristic of BC and MWCNT PM. There was no correlation with total PAH content of the soot PM. Cytokine release (IL-6, IL-8) was detected for the Ag, Fe2 O3, asbestos, BC and the MWCNT PM. Reactive oxygen species (ROS) production was also detected for Ag, Fe2 O3, ZrO2, asbestos, BC, and the MWCNT aggregate PM, as well as the natural gas kitchen burner combustion PM. TEM, FESEM, and optical microscopy examination of these nanomaterials illustrate the wide range in PM morphologies and crystallinities as well as cell morphologies. Taken together, these results illustrate proinflammatory and related respiratory health issues in relation to environmental nanoparticulates.

Microstructures and nanostructures for environmental carbon nanotubes and nanoparticulate soots.

This paper examines the microstructures and nanostructures for natural (mined) chrysotile asbestos nanotubes (Mg3 Si2O5 (OH)4) in comparison with commercial multiwall carbon nanotubes (MWCNTs), utilizing scanning and transmission electron microscopy (SEM and TEM). Black carbon (BC) and a variety of specific soot particulate (aggregate) microstructures and nanostructures are also examined comparatively by SEM and TEM. A range of MWCNTs collected in the environment (both indoor and outdoor) are also examined and shown to be similar to some commercial MWCNTs but to exhibit a diversity of microstructures and nanostructures, including aggregation with other multiconcentric fullerenic nanoparticles. MWCNTs formed in the environment nucleate from special hemispherical graphene "caps" and there is evidence for preferential or energetically favorable chiralities, tube growth, and closing. The multiconcentric graphene tubes ( approximately 5 to 50 nm diameter) differentiate themselves from multiconcentric fullerenic nanoparticles and especially turbostratic BC and carbonaceous soot nanospherules ( approximately 8 to 80 nm diameter) because the latter are composed of curved graphene fragments intermixed or intercalated with polycyclic aromatic hydrocarbon (PAH) isomers of varying molecular weights and mass concentrations; depending upon combustion conditions and sources. The functionalizing of these nanostructures and photoxidation and related photothermal phenomena, as these may influence the cytotoxicities of these nanoparticulate aggregates, will also be discussed in the context of nanostructures and nanostructure phenomena, and implications for respiratory health.

Cytotoxic effects of aggregated nanomaterials.

This study deals with cytotoxicity assays performed on an array of commercially manufactured inorganic nanoparticulate materials, including Ag, TiO(2), Fe(2)O(3), Al(2)O(3), ZrO(2), Si(3)N(4), naturally occurring mineral chrysotile asbestos and carbonaceous nanoparticulate materials such as multiwall carbon nanotube aggregates and black carbon aggregates. The nanomaterials were characterized by TEM, as the primary particles, aggregates or long fiber dimensions ranged from 2nm to 20microm. Cytotoxicological assays of these nanomaterials were performed utilizing a murine alveolar macrophage cell line and human macrophage and epithelial lung cell lines as comparators. The nanoparticulate materials exhibited varying degrees of cytoxicity for all cell lines and the general trends were similar for both the murine and human macrophage cell lines. These findings suggest that representative cytotoxic responses for humans might be obtained by nanoparticulate exposures to simple murine macrophage cell line assays. Moreover, these results illustrate the utility in performing rapid in vitro assays for cytotoxicity assessments of nanoparticulate materials as a general inquiry of potential respiratory health risks in humans.

Combustion-generated nanoparticulates in the El Paso, TX, USA / Juarez, Mexico Metroplex: their comparative characterization and potential for adverse health effects.

In this paper we report on the collection of fine (PM1) and ultrafine (PM0.1), or nanoparticulate, carbonaceous materials using thermophoretic precipitation onto silicon monoxide/formvar-coated 3 mm grids which were examined in the transmission electron microscope (TEM). We characterize and compare diesel particulate matter (DPM), tire particulate matter (TPM), wood burning particulate matter, and other soot (or black carbons (BC)) along with carbon nanotube and related fullerene nanoparticle aggregates in the outdoor air, as well as carbon nanotube aggregates in the indoor air; and with reference to specific gas combustion sources. These TEM investigations include detailed microstructural and microdiffraction observations and comparisons as they relate to the aggregate morphologies as well as their component (primary) nanoparticles. We have also conducted both clinical surveys regarding asthma incidence and the use of gas cooking stoves as well as random surveys by zip code throughout the city of El Paso. In addition, we report on short term (2 day) and longer term (2 week) in vitro assays for black carbon and a commercial multiwall carbon nanotube aggregate sample using a murine macrophage cell line, which demonstrate significant cytotoxicity; comparable to a chrysotile asbestos nanoparticulate reference. The multi-wall carbon nanotube aggregate material is identical to those collected in the indoor and outdoor air, and may serve as a surrogate. Taken together with the plethora of toxic responses reported for DPM, these findings prompt concerns for airborne carbonaceous nanoparticulates in general. The implications of these preliminary findings and their potential health effects, as well as directions for related studies addressing these complex issues, will also be examined.

Cytotoxicity assessment of some carbon nanotubes and related carbon nanoparticle aggregates and the implications for anthropogenic carbon nanotube aggregates in the environment.

Nanotechnology and nanomaterials have become the new frontier world-wide over the past few years and prospects for the production and novel uses of large quantities of carbon nanotubes in particular are becoming an increasing reality. Correspondingly, the potential health risks for these and other nanoparticulate materials have been of considerable concern. Toxicological studies, while sparse, have been concerned with virtually uncharacterized, single wall carbon nanotubes, and the conclusions have been conflicting and uncertain. In this research we performed viability assays on a murine lung macrophage cell line to assess the comparative cytotoxicity of commercial, single wall carbon nanotubes (ropes) and two different multiwall carbon nanotube samples; utilizing chrysotile asbestos nanotubes and black carbon nanoaggregates as toxicity standards. These nanotube materials were completely characterized by transmission electron microscopy and observed to be aggregates ranging from 1 to 2 microm in mean diameter, with closed ends. The cytotoxicity data indicated a strong concentration relationship and toxicity for all the carbon nanotube materials relative to the asbestos nanotubes and black carbon. A commercial multiwall carbon nanotube aggregate exhibiting this significant cell response was observed to be identical in structure to multiwall carbon nanotube aggregates demonstrated to be ubiquitous in the environment, and especially in indoor environments, where natural gas or propane cooking stoves exist. Correspondingly, preliminary epidemiological data, although sparse, indicate a correlation between asthma incidence or classification, and exposure to gas stoves. These results suggest a number of novel epidemiological and etiological avenues for asthma triggers and related respiratory or other environmental health effects, especially since indoor number concentrations for multiwall carbon nanotube aggregates is at least 10 times the outdoor concentration, and virtually all gas combustion processes are variously effective sources. These results also raise concerns for manufactured carbon nanotube aggregates, and related fullerene nanoparticles.

Carbon nanotubes and other fullerene nanocrystals in domestic propane and natural gas combustion streams.

Carbon nanotubes and other aggregated fullerene-related multi-layer shell structures have been collected in propane and natural gas flame emissions from domestic cooking stoves and observed by transmission electron microscopy. Some aggregated nanoparticles collected on 3 mm electron microscope grids by thermal precipitation were mostly multi-walled nanotubes; many tangled and distorted, and aggregated with other closed-concentric, multi-shell forms. Such clean-burning regimes may be major contributors to complex particulate matter in indoor and outdoor air.

Chemistry and nanoparticulate compositions of a 10,000 year-old ice core melt water.

Particulates extracted from a single section of a 10,000 year-old ice core melt sample exhibited characteristics of contemporary, airborne fine particulates: a majority were microcrystalline particulates and aggregated microcrystals, including some mixtures of microcrystals and carbonaceous matter. Particularly significant were the presence of carbon nanotubes and fullerene nanocrystals composing aggregated particulates reflecting global combustion products similar to contemporary, airborne carbon nanocrystal aggregates. ICP elemental analysis of the melt water showed significant concentrations of Ca, K and especially Na (corresponding to K, NaCl), S, Si, Se, and Zn. Overall, the elemental analysis of the melt water is similar to local tap water. However, lead was absent in the local tap water and only half the concentration of selenium was present in the tap water in contrast to the ice core water. While these observations cannot be generalized, the methodology illustrates the potential to characterize and compare airborne particulate regimes and water chemistries in antiquity.

Characterization of nanostructure phenomena in airborne particulate aggregates and their potential for respiratory health effects.

Airborne aggregates of nanoparticulates were collected on carbon/form-coated, 100-mesh Ni TEM grids in a thermal precipitator and observed in an analytical TEM utilizing a BF-SAED-DF-EDS characterization protocol to identify the nanocrystalline or nanoparticulate components, especially their degree of crystallinity, size, structural/morphologic features, and chemistries. Reference aggregates of TiO2 rutile and anatase as well as Si3N4 nanoparticles were used to establish these characterization protocols, which were applied to several hundred individual particulates: homogeneous aggregates of carbonaceous/diesel particulate matter, complex mixtures of carbonaceous matter, including carbon nanocrystals, and inorganic nanocrystals; and heterogeneous, nanocrystal/nanoparticulate aggregates. Most airborne particulates were aggregates ranging in aerodynamic diameters from a few nanometers to a few microns; containing as few as 2 nanocrystals to several thousand nanocrystals or nanoparticulates such as carbonaceous spherules arranged in complex branched homogeneous aggregates composing diesel exhaust, with spherule diameters ranging from 10 to 30 nm. The potential for ultrafine airborne aggregates to fragment into hundreds or thousands of nanoparticulate components in human airways and act as toxic agents in deep lung tissue is demonstrated.

Evaluation of mechanical and corrosion biocompatibility of TiTa alloys.

As-received and heat-treated Ti40Ta and Ti50Ta alloys were evaluated to determine their corrosion as well as mechanical performances and compared to Ti6A14V, a common material utilized for orthopedic (surgical) implants. Anodic potentiodynamic tests performed in Plasmalyte showed that all samples, except for the Ti50Ta specimen aged at 400 degrees C for 3 h gave a curve similar to that of Ti6A14V. Optical and TEM microscopy was performed to determine as-received and heat-treated microstructures. As-received materials showed an alpha precipitate in an alpha+beta and martensite matrix. Samples that were aged at 400 degrees C increased in the density and the length of the alpha precipitate. Vickers hardness measurements were performed to get an approximation of the tensile strengths. Aged Ti40Ta and Ti50Ta specimens produced the highest tensile values when compared to the Ti6A14V material, representing a 31% and 56% increase for the 3 h samples and an 18% and 58% increase for the 10 h samples. Of all the materials studied the Ti50Ta specimen aged for 10 h exhibited the best biocompatibility showing excellent corrosion resistance combined with the highest tensile strength (1089 MPa and 58% harder/stronger than Ti6A14V).

A study of alternative metal particle structures and mixtures for dental amalgams based on mercury additions.

The perception that mercury in dental amalgam is toxic to the human organism has prompted worldwide efforts by the scientific community to develop alternative amalgam-like materials that utilize little or no mercury. In this investigation, an attempt is made to develop a new dental alloy system by adding liquid mercury to silver-coated Ag4Sn intermetallic particles in lesser amounts than are used in conventional amalgam alloys. An effort to precipitate the important eta-prime (Cu6Sn5) phase was made by adding pure Cu and Sn powders to the alloy formulation during trituration. Tytin a popular Ag-Sn-Cu single-composition, spray-atomized conventional dental alloy was used as the control to obtain baseline data for comparisons of microstructures and mechanical properties. Amalgamation of the coated particles with mercury, with or without the addition of Cu and Sn powders, mostly produced specimens with chemically non-coherent microstructures that were relatively weak in compression. These results were due, in part, to mercury's inability to chemically wet the Ag-coated particles and Cu and Sn powders because of naturally occurring surface oxide films. The strongest specimens tested had silver dendritic coatings, resulting in compression strength values up to 40% of the control's. Their higher strength is attributed to mechanical interlocking at the particle/matrix interfaces.